Monitoring expression of cell associated transgenes /vector core

本项目的目标是：1)开发一种方便、可靠和普遍适用的策略来监测任何载体编码的转基因在体内的表达；2)为所有PPG研究人员提供具有良好特征的逆转录病毒、慢病毒和腺病毒基因治疗载体制剂，以满足他们的个人需求。在任何体内基因治疗研究中，治疗性转基因表达的概况是结果的关键决定因素，但这一信息通常是不可获得的。当前项目的总体假设是，通过测量体液中可溶性、非免疫原性标记多肽的浓度，可以在体内无创地监测治疗性转基因的表达，这些标记多肽是由同一载体结构一致表达的。具体目的是：1)精确定义通过内部核糖体进入位点（IRES）连接表达的载体编码多肽的合成速率之间的一致性；2)确定由双链慢病毒和腺病毒载体制备的含有一致表达- hcg标记多肽可选特征的逆转录病毒、慢病毒或腺病毒载体表达的可溶性标记肽的体液浓度之间的体内基因表达谱和随时间的一致性，以支持其在免疫能力强的大鼠、狗和猪的临床前研究。

The goals of this project/core are: 1) to develop a convenient , reliable and generally applicable strategy to monitor in vivo the expression of any vector encoded transgene; and 2) to supply all PPG investigators with well-characterized preparations of retroviral, lentiviral and adenoviral gene therapy vectors tailored to their individual requirements. The profile of therapeutic transgene expression is a key determinant of the outcome in any in vivo gene therapy study, but this information is usually not available. The overall hypothesis for the current project is that the expression of the therapeutic transgene can be non-invasively monitored in vivo by measuring the concentration in body fluids of a soluble, non- immunogenic marker polypeptide concordantly expressed from the same vector construct. The Specific Aims are: 1) To precisely define the concordance between the rates of synthesis of vector encoded polypeptides whose expression is linked through an internal ribosome entry site (IRES); 2) To define the in vivo gene expression profiles and concordance over time between body fluid concentrations of soluble marker peptides expressed from the bicistronic lentiviral and adenoviral vectors of preparations of retroviral, lentiviral or adenoviral vectors incorporating the optional feature of concordantly expressed beta-hCG marker polypeptide to support their preclinical studies in immunocompetent rats, dogs and pigs.